Bead Array Direct rRNA Capture Assay (rCapA) for Amplification Free Speciation of Mycobacterium Cultures

Mycobacterium cultures, from patients suspected of tuberculosis or nontuberculous mycobacteria (NTM) infection, need to be identified. It is most critical to identify cultures belonging to the Mycobacterium tuberculosis complex, but also important to recognize clinically irrelevant or important NTM to allow appropriate patient management. Identification of M. tuberculosis can be achieved by a simple and cheap lateral flow assay, but identification of other Mycobacterium spp. generally requires more complex molecular methods. Here we demonstrate that a paramagnetic liquid bead array method can be used to capture mycobacterial rRNA in crude lysates of positive cultures and use a robust reader to identify the species in a direct and sensitive manner. We developed an array composed of paramagnetic beads coupled to oligonucleotides to capture 16 rRNA from eight specific Mycobacterium species and a single secondary biotinilated reporter probe to allow the captured rRNA to be detected. A ninth less specific bead and its associated reporter probe, designed to capture 23S rRNA from mycobacteria and related genera, is included as an internal control to confirm the presence of bacterial rRNA from a GC rich Gram variable genera. Using this rRNA capture assay (rCapA) with the array developed we were already able to confirm the presence of members of the M. tuberculosis complex and to discriminate a range of NTM species. This approach is not based on DNA amplification and therefore does not require precautions to avoid amplicon contamination. Moreover, the new generation of stable and cost effective liquid bead readers provides the necessary multiplexing potential to develop a robust and highly discriminatory assay

Caregiver Burdens and Preventive Dental Care for Children with Autism Spectrum Disorder, Developmental Disability and/or Mental Health Conditions: National Survey of CSHCN, 2009–2010

OBJECTIVE: The purpose of this study is to examine the burdens of caregivers on one perception of the need and receipt of preventive dental care for a subset of children with special health care needs—children with Autism Spectrum disorder, developmental disability and/or mental health conditions (CASD/DD/MHC). METHODS: The authors used the 2009–2010 National Survey of CSHCN. The survey included questions addressing preventive dental care and caregivers’ financial, employment, and time-related burdens. The associations of these burdens on perceptions and receipt of preventive dental care use were analyzed with bivariate Chi square analyses and multinomial logistic regressions for CASD/DD/MHC (N=16,323). RESULTS: Overall, 16.3% of CASD/DD/MHC had an unmet preventive dental care need. There were 40.0% of caregivers who reported financial burden, 20.3% who reported employment burden, and 10.8% who reported time burden. A higher percentage of caregivers with financial burden, employment burden, and time-related burden reported that their CASD/DD/MHC did not receive needed preventive dental care (14.1 %, 16.5%, 17.7% respectively) compared to caregivers without financial, employment, or time burdens (9.0%, 9.6%, 11.0% respectively). Caregivers with financial burden (adjusted multinomial odds ratio, 1.38 [95%CI: 1.02, 1.86]) and employment burden (adjusted multinomial odds ratio, 1.45 [95%CI: 1.02, 2.06]) were more likely to report that their child did not receive preventive dental care despite perceived need compared to caregivers without financial or employment burdens. CONCLUSIONS FOR PRACTICE: Unmet needs for preventive dental care were associated with employment and financial burdens of the caregivers of CASD/DD/MHC